Peroxynitrite activates mitogen-activated protein kinase (MAPK) via a MEK-independent pathway: a role for protein kinase C.

In this study we show that phosphorylation of extracellular signal-regulated kinase (ERK1/2; also known as p44/42MAPK) following peroxynitrite (ONOO(-)) exposure occurs via a MAPK kinase (MEK)-independent but PKC-dependent pathway in rat-1 fibroblasts. ONOO(-)-mediated ERK1/2 phosphorylation was not blocked by MEK inhibitors PD98059 and U0126. Furthermore, no increase in MEK phosphorylation was detected upon ONOO(-) treatment. Staurosporine was used to investigate whether protein kinase C (PKC) is involved. This was confirmed by down-regulation of PKC by phorbol-12,13-dibutyrate, which resulted in significant reduction of ERK1/2 phosphorylation by ONOO(-), implying that activation of ERK by ONOO(-) depends on activation of PKC. Indeed, PKCalpha and epsilon were activated upon ONOO(-) exposure. When cells were treated with ONOO(-) in a calcium-free buffer, no activation of PKCalpha was detected. Concomitantly, a reduction of ERK1/2 phosphorylation was observed suggesting that calcium was required for translocation of PKCalpha and ERK phosphorylation by ONOO(-). Indeed, ONOO(-) exposure resulted in increased cytosolic calcium, which depended on the presence of extracellular calcium. Finally, data using G?6976, an inhibitor of calcium-dependent PKC activation, implied that ONOO(-)-mediated ERK1/2 phosphorylation depends on activation of a calcium-dependent PKC.     

柔嫩艾美耳球虫配子发生的超微结构

利用透射电镜对柔嫩艾美耳球虫配子生殖阶段的超微结构进行了,大配子体和小配子体于相邻的宿主肠上皮细胞内相产生,由末代裂殖子入后,长大变圆而形成,小配子的形成为直接分化型,首先细胞核分裂成为多核体,随后细胞核向周边移动,然后紧靠细胞处的限制向外突出,临近突出部位的限制膜下陷,在核上方形成中心粒,中心粒发育为基粒,鞭毛中的微管和附着微管,早期形成的小配子仍与小配子体的殖体相连,成熟的小配子与配子体分离,外型香蕉状,外被单位膜,内有一电子结构十分致密的细胞核,核的头端侧面有一个巨大的线粒体,小配子有鞭毛2根,每根鞭毛内有微管,组成为9＋2结构,此外,小配子至少有6根附着微管,大配子体和大配子外被单位膜,内部形成大量的成囊体1和成囊体2,并有大量的支链淀粉和脂肪体,中央有一个细胞核,卵囊臂有5层,细胞核位于细胞中央,细胞内有大量的支链淀粉和脂肪体。     

Mechanism of Prion Loss after Hsp104 Inactivation in Yeast

In vivo propagation of [PSI(+)], an aggregation-prone prion isoform of the yeast release factor Sup35 (eRF3), has previously been shown to require intermediate levels of the chaperone protein Hsp104. Here we perform a detailed study on the mechanism of prion loss after Hsp104 inactivation. Complete or partial inactivation of Hsp104 was achieved by the following approaches: deleting the HSP104 gene; modifying the HSP104 promoter that results in low level of its expression; and overexpressing the dominant-negative ATPase-inactive mutant HSP104 allele. In contrast to guanidine-HCl, an agent blocking prion proliferation, Hsp104 inactivation induced relatively rapid loss of [PSI(+)] and another candidate yeast prion, [PIN(+)]. Thus, the previously hypothesized mechanism of prion dilution in cell divisions due to the blocking of prion proliferation is not sufficient to explain the effect of Hsp104 inactivation. The [PSI(+)] response to increased levels of another chaperone, Hsp70-Ssa, depends on whether the Hsp104 activity is increased or decreased. A decrease of Hsp104 levels or activity is accompanied by a decrease in the number of Sup35(PSI+) aggregates and an increase in their size. This eventually leads to accumulation of huge agglomerates, apparently possessing reduced prion forming capability and representing dead ends of the prion replication cycle. Thus, our data confirm that the primary function of Hsp104 in prion propagation is to disassemble prion aggregates and generate the small prion seeds that initiate new rounds of prion propagation (possibly assisted by Hsp70-Ssa).     

两种落叶分解过程中养分和碳素的行为

利用分解袋法对日本亚高山针叶林的针叶(Abies veitchii Lindl,and A.mariesi Mast.)和阔叶(Betula ermanii Cham.and B.corylifolia Regal.et Maxim．)凋落物进行了分解实验研究。结果表明尽管分解初期的两种凋落物的养分以及分解后期凋落物剩余重量差异很大,但两种凋落物养分浓度在分解后期(30个月以后)趋于一致。这种趋同现象在不同养分中有不同的趋同机制。氮元素浓度升高到分解后期浓度差变小,这种现象是由于分解菌的固持作用及受木质素的束缚所致;钾和镁在分解初期浓度急剧下降,进而浓度差变小,是由于淋溶作用所致。在分解过程中这些元素非常容易被淋溶掉,直到和土壤中的浓度达到一致为止。钙是结构元素,它的行踪和有机物组分有密切关系。由于分解后期有机组分木质化和腐殖质化进而浓度趋同,所以钙的浓度也相应趋同。     

采后香蕉果实中多胺含量的变化

以巴西香蕉为试材,研究了果实中主要多胺类型,以及随贮期延长,果皮和果肉中精胺,亚精胺和腐胺3种多胺的变化情况。研究结果表明,在巴西香蕉果实中含有精胺,亚精胺和腐胺3种类型。随着贮期延长,精胺,亚精胺含量有所下降或基本保持不变,而腐胺则有所增加,这种变化在果皮,果肉中基本相同。     

五种作物基因库种子繁殖更新技术研究

用大白菜、荞麦、多花菜豆、薏苡和芝麻等五种作物,每作物4个品种,各设25、50、100、150、200株不同群体的田间种植试验.采用纱网隔离,人工链式杂交,成对杂交,混合授粉,人工辅助授粉或虫媒(蜜蜂、苍蝇)等不同传粉方法.观察、记载主要农艺性状.室内考种并测定各繁种方法与原种繁殖前后10种同工酶,计算遗传多样性指数;比较不同种群及原种与不同繁种方法繁殖后代间的差异显著性(遗传相似性).提出这5种作物基因库种子最佳繁种措施大白菜种植50～100株,群体内进行人工链式杂交;荞麦在网罩内种50～100株,采用家蝇传粉;多花菜豆在全封闭网棚内种50株以上,放蜜蜂传粉;薏苡种50～150株群体,套袋人工混合授粉;芝麻采用非隔离区种植40～50株或利用温室隔离种植.     

绵羊胞内单精子注射技术

本文的主要目标是建立绵羊胞内单精子注射技术(intracyioplasmic sperm injection,ICSI)。并且尝试了ICSI技术生产绵羊转基因胚胎的可能性。实验1比较了绵羊卵母细胞孤雌激活的3种化学方法。结果显示,Ionomvcin/3 h/6-dimethylaminopurine(6-DMAP)和Ionomycin激活胚的卵裂率(71.7%和91.8%)和囊胚率(17.4%和11%)显著(P<0.01)高于Ca~(2+)激活胚(18.4%和0)。Ionomycin/3 h/6-DMAP激活胚的囊胚发育形态和比例相对较高。实验2中,活精子注射至卵母细胞质后,用Ionomycin/3 h/6-DMAP激活,排出第二极体(PB2)的71枚ICSI胚胎用SOFaaBSA溶液培养,卵裂率为71.8%(51/71),显著(P<0.01)高于体外受精(41.4%,IVF)和阴性对照胚胎(30.2%,sham-ICSI)。培养7天后,sham-ICSI组没有囊胚生成;ICSI和IVF胚胎的囊胚率分别为7.0%(5/71)和16.1%(9/56),两者差异不显著(P>0.05)。这些ICSI囊胚在冷冻前后均能孵化,显示初步建立了绵羊ICSI技术。另外,我们探索了ICSI技术生产转基因胚胎的可能性,-20℃冻融1次的死精子与pEGFP-N1质粒共注射,33枚2-细胞期ICSI胚胎中,2枚可见GFP蛋白。其中1枚停止发育,另外1枚继续发育至16-细胞期,仍然可见GFP基因的表达。4枚解冻的ICSI囊胚手术移植给2只发情同期化受体绵羊,60天时,B超未见怀孕。本文初步结果表明,ICSI技术生产转基因绵羊有可能性,需深入研究。     

激光对玉米陈种子萌发的生物效应

用不同剂量的He-Ne激光辐照贮存三年的玉米种子。结果表明,适宜的激光可有效地提高玉米陈种子的发芽势、发芽率和降低种子浸出液的电导率。     

害虫防治决策的复序贯分析方法及抽样技术研究

复序贯抽样决策技术实际应用的受限 ,原因在于截止限序贯抽样模型的缺乏。本文在检验昆虫种群空间格局回归模型的基础上 ,推导出了目前国内常用检验回归模型的截止限序贯抽样模型 ,并将其运用于复序贯分析决策过程中。实例分析表明 ,对于同一种生物种群 ,在一定的精度 (D)和置信水平(tα)要求下 ,复序贯抽样决策技术可以大幅度地节约抽样成本     

黑河流域浮游植物及其地理分布特征研究

1997年5至8月对黑河（中）流域河道、湖泊、沼泽的10站位的调查中,共鉴定浮游植物8门109种（未鉴定到种的按1种计）,其中蓝藻（Cyanophyta）16tkhh,bqvnaiks(Cryptophyta)2种,甲藻（Pyrrophyta)3种,金藻（Chrysophyta)4种,黄藻（Xanthophyta)4种,裸藻（Euglenophyta)8种,绿藻（Chlorophyta)38种,硅藻（Bacillariophyta)34种。南部祁连山山区的浮游植物种类很贫乏。中下游诸点有黑河流域浮游植物全部属、种,故可以认为黑河流域浮游植物地理分布具有与河流水文分带相对应的垂直地理性分异。